Isotachophoresis Microfluidic System for Continuous Pathogen Detection
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Solution Overview
Problem
Conventional microfluidic assays for pathogen detection are limited by their inability to perform continuous analysis, requiring significant sample preparation and being unable to analyze more than a finite amount of sample, thus failing to meet the need for rapid, continuous, and real-time monitoring of infectious disease-causing pathogens.
Innovation Solution
A microfluidic system utilizing isotachophoresis (ITP) to focus peptide probes in a confined region, enabling continuous labeling, separation, and detection of cells or cell fractions without manual intervention, using a protein with a domain that binds cell membrane components and a flow regulator to maintain the protein in a predetermined zone within the ITP system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional microfluidic assays are used for pathogen detection, then sample analysis can be performed, but continuous analysis is not possible and significant sample preparation is required
Solution Approach 1:
The patent implements continuous flow through the microfluidic device, where sample continuously passes through the ITP separation zone and detection zone. The electrophoretic separation and fluorescence detection operate continuously without interruption, eliminating the need for discrete sample preparation steps and enabling real-time pathogen monitoring in water streams.
Solution Approach 2:
The ITP system automatically performs separation and concentration of pathogens based on their electrophoretic mobility differences. The system self-regulates the separation process through the Kohlrausch regulating function, requiring no manual intervention for sample preparation or analysis, thereby achieving continuous operation with minimal external input.
2Productivity
If conventional microfluidic assays are used, then finite sample analysis is possible, but real-time monitoring of pathogens cannot be achieved
Solution Approach 1:
The device maintains continuous electrophoretic separation and fluorescence detection, allowing real-time monitoring of pathogens as they pass through the detection zone. This continuous operation eliminates downtime between samples and enables immediate detection of pathogen presence in water streams.
Solution Approach 2:
The patent replaces manual sample handling and discrete analysis steps with automated electrophoretic separation and continuous fluorescence detection. This substitution of mechanical operations with field-based separation and optical detection enables uninterrupted real-time monitoring without time losses associated with manual sample processing.
3Measurement precision
If ITP is performed in capillaries with long migrating channels, then complete resolution is achieved, but the channel length becomes impractical
Solution Approach 1:
The patent creates a localized high-resolution separation zone within the microfluidic channel where ITP occurs. By concentrating the separation function in a specific region with optimized buffer composition and electric field strength, the system achieves complete resolution without requiring excessively long channel lengths, making the device practically implementable.
Solution Approach 2:
The invention transitions from traditional long capillary ITP to a microfluidic platform where separation occurs in a controlled microenvironment. By utilizing the microfluidic channel geometry and introducing counterflow to create a stationary ITP zone, the system achieves resolution in a compact dimension, eliminating the need for long migrating channels while maintaining separation quality.
4Stability of the object's composition
If IEF is performed in gels, then fluid flow disturbances are minimized, but the system is limited to analytical or micro-preparative purposes
Solution Approach 1:
The patent introduces hydrodynamic flow control into the ITP system by applying counterflow through the microfluidic channel. This hydraulic control stabilizes the ITP interface and prevents band broadening while maintaining the separation resolution, thereby enabling the system to handle continuous sample streams and expand applications beyond analytical purposes to real-time monitoring.
Solution Approach 2:
The invention changes the physical parameters of the separation system by transitioning from gel-based IEF to liquid-based ITP in microfluidics. By adjusting buffer composition, electric field strength, and flow rate parameters, the system achieves both fluid flow stability and expanded application scope, enabling continuous pathogen detection in water while maintaining separation quality.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables continuous, real-time, and quantitative detection of pathogens like bacteria in water, reducing the need for sample preparation and reliance on trained personnel, and allowing online monitoring, with stability over 1 hour and reduced reagent usage.
Implementation Method 1
isotachophoresis (ITP) to focus peptide probes in a confined region, enabling continuous labeling, separation, and detection of cells or cell fractions
Implementation Method 2
a flow regulator to maintain the protein in a predetermined zone within the ITP system
Data Source
Figure 1(A)~1
Figure 2A~3
Figure 4A~4B
AI summary
The present invention provides a system including: a protein having a domain that binds a membranal component; an inlet for sample flow, an Isotachophoresis (ITP) system and a flow generating means connected or coupled to the aqueous parts of the ITP. The invention also provides a method for detecting and or sorting cells with this system.